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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1989 Mar;27(3):377–384. doi: 10.1128/jcm.27.3.377-384.1989

Production of antibody to and cellular localization of erythrocyte-sensitizing substance from Rickettsia rickettsii.

K E Hechemy 1, J Fox 1, W A Samsonoff 1, R Anacker 1, D Silverman 1, C Eisemann 1, I S Green 1
PMCID: PMC267325  PMID: 2497137

Abstract

Antibodies to Rickettsia rickettsii erythrocyte-sensitizing substance (ESS) were raised in rabbits by using a derivatized ESS. The resulting antibodies reacted with R. rickettsii and cross-reacted with Rickettsia conorii, a member of the spotted fever group rickettsiae, but did not react with Rickettsia typhi, a member of the typhus group rickettsiae, Legionella bozemanii, or Proteus vulgaris OX19 or OX2. Immunoblot analysis indicated that ESS was present in more than one fraction and that the major haptenic fraction was proteinase resistant. Immunoelectron microscopy indicated that the antibodies to R. rickettsii were specific to components located on the cell surface and intracellularly to components between the cell wall and cytoplasmic membrane.

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Selected References

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  1. Anacker R. L., Gerloff R. K., Thomas L. A., Mann R. E., Brown W. R., Bickel W. D. Purification of Rickettsia rickettsi by density-gradient zonal centrifugation. Can J Microbiol. 1974 Nov;20(11):1523–1527. doi: 10.1139/m74-238. [DOI] [PubMed] [Google Scholar]
  2. Anacker R. L., List R. H., Mann R. E., Hayes S. F., Thomas L. A. Characterization of monoclonal antibodies protecting mice against Rickettsia rickettsii. J Infect Dis. 1985 Jun;151(6):1052–1060. doi: 10.1093/infdis/151.6.1052. [DOI] [PubMed] [Google Scholar]
  3. Anacker R. L., Philip R. N., Casper E., Todd W. J., Mann R. E., Johnston M. R., Nauck C. J. Biological properties of rabbit antibodies to a surface antigen of Rickettsia rickettsii. Infect Immun. 1983 Apr;40(1):292–298. doi: 10.1128/iai.40.1.292-298.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Anacker R. L., Philip R. N., Thomas L. A., Casper E. A. Indirect hemagglutination test for detection of antibody to Rickettsia rickettsii in sera from humans and common laboratory animals. J Clin Microbiol. 1979 Nov;10(5):677–684. doi: 10.1128/jcm.10.5.677-684.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. CHANG R. S., MURRAY E. S., SNYDER J. C. Erythrocyte-sensitizing substances from Rickettsiae of the Rocky Mountain spotted fever group. J Immunol. 1954 Jul;73(1):8–15. [PubMed] [Google Scholar]
  6. CHANG S. M. A serologically-active erythrocyte-sensitizing substance from typhus rickettsiae. I. Isolation and titration. J Immunol. 1953 Mar;70(3):212–214. [PubMed] [Google Scholar]
  7. CHANG S. M., SNYDER J. C., MURRAY E. S. A serologically active erythrocyte sensitizing substance from typhus rickettsiae. II. Serological properties. J Immunol. 1953 Mar;70(3):215–221. [PubMed] [Google Scholar]
  8. Craig S., Goodchild D. J. Post-embedding immunolabelling. Some effects of tissue preparation on the antigenicity of plant proteins. Eur J Cell Biol. 1982 Oct;28(2):251–256. [PubMed] [Google Scholar]
  9. Eldred W. D., Zucker C., Karten H. J., Yazulla S. Comparison of fixation and penetration enhancement techniques for use in ultrastructural immunocytochemistry. J Histochem Cytochem. 1983 Feb;31(2):285–292. doi: 10.1177/31.2.6339606. [DOI] [PubMed] [Google Scholar]
  10. Erlanger B. F. The preparation of antigenic hapten-carrier conjugates: a survey. Methods Enzymol. 1980;70(A):85–104. doi: 10.1016/s0076-6879(80)70043-5. [DOI] [PubMed] [Google Scholar]
  11. Hawkes R., Niday E., Gordon J. A dot-immunobinding assay for monoclonal and other antibodies. Anal Biochem. 1982 Jan 1;119(1):142–147. doi: 10.1016/0003-2697(82)90677-7. [DOI] [PubMed] [Google Scholar]
  12. Hechemy K. E., Anacker R. L., Carlo N. L., Fox J. A., Gaafar H. A. Absorption of Rickettsia rickettsii antibodies by Rickettsia rickettsii antigens in four diagnostic tests. J Clin Microbiol. 1983 Mar;17(3):445–449. doi: 10.1128/jcm.17.3.445-449.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hechemy K. E., Anacker R. L. Coating of polymeric surfaces for immunoassay by a forced adsorption technique. J Immunoassay. 1983;4(2):147–157. doi: 10.1080/15321818308057007. [DOI] [PubMed] [Google Scholar]
  14. Hechemy K. E., Anacker R. L., Philip R. N., Kleeman K. T., MacCormack J. N., Sasowski S. J., Michaelson E. E. Detection of Rocky Mountain spotted fever antibodies by a latex agglutination test. J Clin Microbiol. 1980 Aug;12(2):144–150. doi: 10.1128/jcm.12.2.144-150.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hechemy K. E., Osterman J. V., Eisemann C. S., Elliott L. B., Sasowski S. J. Detection of typhus antibodies by latex agglutination. J Clin Microbiol. 1981 Jan;13(1):214–216. doi: 10.1128/jcm.13.1.214-216.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hitchcock P. J. Analyses of gonococcal lipopolysaccharide in whole-cell lysates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis: stable association of lipopolysaccharide with the major outer membrane protein (protein I) of Neisseria gonorrhoeae. Infect Immun. 1984 Oct;46(1):202–212. doi: 10.1128/iai.46.1.202-212.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kitagawa T., Aikawa T. Enzyme coupled immunoassay of insulin using a novel coupling reagent. J Biochem. 1976 Jan;79(1):233–236. doi: 10.1093/oxfordjournals.jbchem.a131053. [DOI] [PubMed] [Google Scholar]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Liu F. T., Zinnecker M., Hamaoka T., Katz D. H. New procedures for preparation and isolation of conjugates of proteins and a synthetic copolymer of D-amino acids and immunochemical characterization of such conjugates. Biochemistry. 1979 Feb 20;18(4):690–693. doi: 10.1021/bi00571a022. [DOI] [PubMed] [Google Scholar]
  20. Murphy J. R., Fiset P., Snyder L. B., Wisseman C. L., Jr Antibody to Rickettsia mooseri erythrocyte-sensitizing substance. Infect Immun. 1979 Jun;24(3):962–964. doi: 10.1128/iai.24.3.962-964.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. NETER E., WESTPHAL O., LUDERITZ O., GORZYNSKI E. A., EICHENBERGER E. Studies of enterobacterial lipopolysaccharides; effects of heat and chemicals on erythrocyte-modifying, antigenic, toxic and pyrogenic properties. J Immunol. 1956 May;76(5):377–385. [PubMed] [Google Scholar]
  22. O'Sullivan M. J., Gnemmi E., Morris D., Chieregatti G., Simmonds A. D., Simmons M., Bridges J. W., Marks V. Comparison of two methods of preparing enzyme-antibody conjugates: application of these conjugates for enzyme immunoassay. Anal Biochem. 1979 Nov 15;100(1):100–108. doi: 10.1016/0003-2697(79)90117-9. [DOI] [PubMed] [Google Scholar]
  23. Osterman J. V., Eisemann C. S. Rickettsial indirect hemagglutination test: isolation of erythrocyte-sensitizing substance. J Clin Microbiol. 1978 Aug;8(2):189–196. doi: 10.1128/jcm.8.2.189-196.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Philip R. N., Casper E. A., MacCormack J. N., Sexton D., Thomas L. A., Anacker R. L., Burgdorfer W., Vick S. A comparison of serologic methods for diagnosis of Rocky Mountain spotted fever. Am J Epidemiol. 1977 Jan;105(1):56–67. doi: 10.1093/oxfordjournals.aje.a112356. [DOI] [PubMed] [Google Scholar]
  25. Philip R. N., Casper E. A., Ormsbee R. A., Peacock M. G., Burgdorfer W. Microimmunofluorescence test for the serological study of rocky mountain spotted fever and typhus. J Clin Microbiol. 1976 Jan;3(1):51–61. doi: 10.1128/jcm.3.1.51-61.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. SCHAECHTER M., TOUSIMIS A. J., COHN Z. A., ROSEN H., CAMPBELL J., HAHN F. E. Morphological, chemical, and serological studies of the cell walls of Rickettsia mooseri. J Bacteriol. 1957 Dec;74(6):822–829. doi: 10.1128/jb.74.6.822-829.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. STOENNER H. G., LACKMAN D. B., BELL E. J. Factors affecting the growth of rickettsias of the spotted fever group in fertile hens' eggs. J Infect Dis. 1962 Mar-Apr;110:121–128. doi: 10.1093/infdis/110.2.121. [DOI] [PubMed] [Google Scholar]
  28. Shirai A., Dietel J. W., Osterman J. V. Indirect hemagglutination test for human antibody to typhus and spotted fever group rickettsiae. J Clin Microbiol. 1975 Nov;2(5):430–437. doi: 10.1128/jcm.2.5.430-437.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Silverman D. J., Wisseman C. L., Jr, Waddell A. D., Jones M. External layers of Rickettsia prowazekii and Rickettsia rickettsii: occurrence of a slime layer. Infect Immun. 1978 Oct;22(1):233–246. doi: 10.1128/iai.22.1.233-246.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sompolinsky D., Boldur I., Goldwasser R. A., Kahana H., Kazak R., Keysary A., Pik A. Serological cross-reactions between Rickettsia typhi, Proteus vulgaris OX19, and Legionella bozemanii in a series of febrile patients. Isr J Med Sci. 1986 Oct;22(10):745–752. [PubMed] [Google Scholar]
  31. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weber K., Rathke P. C., Osborn M. Cytoplasmic microtubular images in glutaraldehyde-fixed tissue culture cells by electron microscopy and by immunofluorescence microscopy. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1820–1824. doi: 10.1073/pnas.75.4.1820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Weiss E., Coolbaugh J. C., Williams J. C. Separation of viable Rickettsia typhi from yolk sac and L cell host components by renografin density gradient centrifugation. Appl Microbiol. 1975 Sep;30(3):456–463. doi: 10.1128/am.30.3.456-463.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wood W. H., Jr, Wisseman C. L., Jr Studies of Rickettsia mooseri cell walls. II. Immunologic properties. J Immunol. 1967 Jun;98(6):1224–1230. [PubMed] [Google Scholar]

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